LOCUS       sequence-338305-        5175 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express human Rab5b in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5175)
  AUTHORS   Wu H, Voeltz GK
  TITLE     Reticulon-3 Promotes Endosome Maturation at ER Membrane Contact 
            Sites.
  JOURNAL   Dev Cell. 2021 Jan 11;56(1):52-66.e7. doi: 
            10.1016/j.devcel.2020.12.014.
  PUBMED    33434526
REFERENCE   2  (bases 1 to 5175)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.devcel.2020.12"; journalName: "Dev Cell"; date: 
            "2021-01-11- 11"; volume: "56"; issue: "1"; pages: "52-66.e7"
COMMENT     Sequence Label: SNAP-Rab5
FEATURES             Location/Qualifiers
     source          1..5175
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             613..1158
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     polyA_signal    1963..2084
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2000..2019)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2054..2073
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2091..2546)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2228..2249)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2440..2459
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2573..2677
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2679..3036
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2679..2870
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2702..2722)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2887..3022
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3071..3865
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3071..3865
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3125..3144)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3735..3754
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4053..4072)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4097..4144
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4473..5061
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4962..4981
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggacaa agactgcgaa atgaagcgca ccaccctgga tagccctctg
      661 ggcaagctgg aactgtctgg gtgcgaacag ggcctgcacc gtatcatctt cctgggcaaa
      721 ggaacatctg ccgccgacgc cgtggaagtg cctgccccag ccgccgtgct gggcggacca
      781 gagccactga tgcaggccac cgcctggctc aacgcctact ttcaccagcc tgaggccatc
      841 gaggagttcc ctgtgccagc cctgcaccac ccagtgttcc agcaggagag ctttacccgc
      901 caggtgctgt ggaaactgct gaaagtggtg aagttcggag aggtcatcag ctacagccac
      961 ctggccgccc tggccggcaa tcccgccgcc accgccgccg tgaaaaccgc cctgagcgga
     1021 aatcccgtgc ccattctgat cccctgccac cgggtggtgc agggcgacct ggacgtgggg
     1081 ggctacgagg gcgggctcgc cgtgaaagag tggctgctgg cccacgaggg ccacagactg
     1141 ggcaagcctg ggctgggtag cggtagcggt ggcagctctc gagctatgac tagcagaagc
     1201 acagctaggc ccaatgggca accccaggcc agcaaaattt gccagttcaa attggtcctg
     1261 ctgggagaat ctgcagtggg aaagtcaagc ctggtattac gttttgtcaa agggcagttc
     1321 catgagtacc aggagagcac cattggagcg gccttcctca cccagtccgt ttgtctagat
     1381 gacacaacag tgaagtttga gatctgggac acagctgggc aggagcgata tcacagctta
     1441 gcccccatgt actacagggg tgcccaagct gcaatcgtgg tttacgacat tactaatcag
     1501 gaaacctttg cccgagcaaa gacatgggtg aaggaactac agcgacaggc cagtcctagc
     1561 atcgttattg ccctggcagg gaacaaagct gacctggcca acaaacgtat ggtggagtat
     1621 gaagaggccc aggcatatgc agatgacaac agcttattgt tcatggagac ttcagccaag
     1681 acagctatga acgtgaatga tctcttcctg gcaatagcta agaagttgcc aaagagtgaa
     1741 ccccagaatc tgggaggtgc agcaggccga agccggggtg tggatctcca tgaacagtcc
     1801 cagcagaaca agagccagtg ttgtagcaac tgaggatcca ccggatctag ataactgatc
     1861 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     1921 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     1981 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     2041 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaacgcgt aaattgtaag
     2101 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     2161 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     2221 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     2281 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     2341 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     2401 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     2461 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     2521 gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac
     2581 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     2641 ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg
     2701 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     2761 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     2821 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     2881 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     2941 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     3001 tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg
     3061 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     3121 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     3181 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     3241 gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     3301 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     3361 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     3421 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     3481 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     3541 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag
     3601 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     3661 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     3721 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     3781 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     3841 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     3901 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     3961 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     4021 gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa
     4081 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt
     4141 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
     4201 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt
     4261 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac
     4321 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     4381 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     4441 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     4501 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     4561 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     4621 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     4681 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     4741 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     4801 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     4861 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     4921 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     4981 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     5041 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     5101 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     5161 attaccgcca tgcat
//
